title: Fluorogenic α-tocopherol analogue for monitoring the antioxidant status within the inner mitochondrial membrane of live cells.
abstract: We report here the preparation of a lipophilic fluorogenic antioxidant (Mito-Bodipy-TOH) that targets the inner mitochondrial lipid membrane (IMM) and is sensitive to the presence of lipid peroxyl radicals, effective chain carriers in the lipid chain autoxidation. Mito-Bodipy-TOH enables monitoring of the antioxidant status, i.e., the antioxidant load and ability to prevent lipid chain autoxidation, within the inner mitochondrial membrane of live cells. The new probe consists of 3 segments: a receptor, a reporter, and a mitochondria-targeting element, constructed, respectively, from an α-tocopherol-like chromanol moiety, a BODIPY fluorophore, and a triphenylphosphonium cation (TPP). The chromanol moiety ensures reactivity akin to that of α-tocopherol, the most potent naturally occurring lipid soluble antioxidant, while the BODIPY fluorophore and TPP ensure partitioning within the inner mitochondrial membrane. Mechanistic studies conducted either in homogeneous solution or in liposomes and in the presence of free radical initiators show that the antioxidant activity of Mito-Bodipy-TOH is on par with that of α-tocopherol. Studies conducted on live fibroblast cells further show the antioxidant depletion in the presence of methyl viologen (paraquat), a known agent of oxidative stress and source of superoxide radical anion (and indirectly, a causative of lipid peroxidation) within the mitochondria matrix. We recorded a ca. 8-fold emission enhancement with Mito-Bodipy-TOH in cells stressed with methyl viologen, whereas no enhancement was observed in control studies with untreated cells. Our findings underscore the potential of the new fluorogenic antioxidant Mito-Bodipy-TOH to study the chemical link between antioxidant load, lipid peroxidation and mitochondrial physiology.
authors.affiliation: Department of Chemistry and Center for Self-Assembled Chemical Structures (CSACS-CRMAA), McGill University , 801 Sherbrooke Street West, Montreal, Quebec H3A 0B8, Canada.
authors.forename: Katerina
authors.initials: K
authors.lastname: Krumova
authors.forename: Lana E
authors.initials: LE
authors.lastname: Greene
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Cosa
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000737:chemistry* / Q000378:metabolism; D001896:Boron Compounds / Q000737:chemistry* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D015227:Lipid Peroxidation; D051379:Mice; D018613:Microscopy, Confocal; D051336:Mitochondrial Membranes / Q000737:chemistry / Q000378:metabolism*; D041681:NIH 3T3 Cells; D010545:Peroxides / Q000378:metabolism; D010720:Phosphines / Q000737:chemistry* / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D024502:alpha-Tocopherol / Q000031:analogs & derivatives* / Q000378:metabolism
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D000975:Antioxidants; D001896:Boron Compounds; D005456:Fluorescent Dyes; D010545:Peroxides; D010720:Phosphines; D017382:Reactive Oxygen Species; C049375:perhydroxyl radical; D024502:alpha-Tocopherol